Production of aspirin-triggered resolvins without the use of aspirin in a dietary omega-3 supplement

ABSTRACT

The present invention includes a composition and method of producing aspirin in situ, the method comprising: identifying a subject in need of aspirin or aspirin-like compounds; and providing the subject with a composition comprising: a source of methyl salicylate, an acetyl donor, and L-Arginine, wherein the composition is effective to produce aspirin-triggered resolvins in the subject without the deleterious effect of the aspirin or aspirin-like compounds in the stomach.

CROSS-REFERENCE TO RELATED APPLICATIONS

This patent application is a divisional of U.S. patent application Ser.No. 15/951,755 filed on Apr. 12, 2018 entitled “Production of AspirinTriggered Resolvins Without the Use of Aspirin in a Dietary Omega-3Supplement,” which claims priority to U.S. provisional patentapplication Ser. No. 62/484,676 filed on Apr. 12, 2017 entitled“Production of Aspirin Triggered Resolvins Without the Use of Aspirin ina Dietary Omega-3 Supplement,” all of which is hereby incorporated byreference in their entirety.

STATEMENT OF FEDERALLY FUNDED RESEARCH

Not applicable.

TECHNICAL FIELD OF THE INVENTION

The present invention relates in general to the field of aspirin, andmore particularly, to a novel compositions and methods for producingaspirin in situ without the deleterious side effects of aspirin in thestomach.

BACKGROUND OF THE INVENTION

Without limiting the scope of the invention, its background is describedin connection with aspirin.

One such patent is U.S. Pat. No. 8,586,073, issued to Drapeau, et al.,entitled “Methods and formulations for administration of resolvinanti-inflammatory compounds”. These inventors are said to teachresolvins and their use as anti-inflammatory compounds. The resolvinscan be administered in a variety of forms, including drug depotscomprising polymers or lipids. The pharmaceutical formulations withresolvins are said to be used to treat a variety of conditions includingacute pain and chronic pain.

Another such patent is U.S. Pat. No. 7,341,840, issued to Serhan, etal., entitled “Methods for identification and uses of anti-inflammatoryreceptors for eicosapentaenoic acid analogs”. These inventors are saidto teach methods for the identification and uses of receptors thatinteract with anti-inflammatory compounds derived from eicosapentaenoicacid (EPA). The receptors are of the G-protein coupled receptor (GPCR)family, and are said to be useful for screening candidate substances foranti-inflammatory activity, especially substances that are analogs ofEPA. Such analogs are termed “resolvins”; and are typically di- andtri-hydroxy EPA analogs.

SUMMARY OF THE INVENTION

In one embodiment, the present invention includes a method of producingaspirin or aspirin-like molecule in situ, the method comprising:identifying a subject in need of aspirin or aspirin-like compounds; andproviding the subject with a composition comprising: a source of methylsalicylate, an acetyl donor, and L-Arginine, wherein the composition iseffective to produce aspirin-triggered resolvins in the subject withoutthe deleterious effect of the aspirin or aspirin-like compounds in thestomach. The skilled artisan will recognize that once in the body, thecompounds of the present invention will undergo a variety of chemicalreactions, under varying reaction conditions, and/or in the presence ofenzymes and other chemical modifying agents that may lead to theformation of aspirin or other aspirin-like compounds, which effect canbe measured in the form of pain killing or other known activities ofaspirin and aspirin-like compounds. In one aspect, the source of methylsalicylate is wintergreen oil or eastern teaberry oil. In anotheraspect, the source of methyl salicylate is a Gaultheria sp., Betula sp.,Spiraea sp. or a Polygala sp. In another aspect, the methyl salicylateis provided in an amount between 10 mg and 60 mg. In another aspect, theacetyl donor is provided in an amount between 30 mg and 300 mg. Inanother aspect, the L-Arginine is provided in an amount between 3 mg and40 mg. In another aspect, the method further comprises packaging thecomposition into a gelcap, tablet, powder, cream, lotion, liquid,softgel, poultice, suppository, or serum. In another aspect, the methodfurther comprises formulating the composition for oral, sublingual,subcutaneous, percutaneous, intramuscular, nasal, intradermal,pulmonary, intraalveolar, intravaginal, intrarectal, intraperitoneal orintravenous administration. In another aspect, the compositiondemonstrates increase potency and reduced off-target effects whencompared to aspirin, aspirin-like products, or taken separately. Inanother aspect, the acetyl donor is d,l-alpha-Tocopherol Acetate,Vitamin E, or an acetylated vitamin. In another aspect, the methodfurther comprises adding an Omega-3 dietary supplement to thecomposition. In another aspect, the method further comprises addingeicosapentaenoic acid (EPA) to the composition. In another aspect, themethod further comprises adding docosahexaenoic acid (DHA) to thecomposition. In another aspect, the composition comprises an emulsifierselected from beeswax, ammonium lauryl sulfate, sodium laureth sulfate,sodium oleyl succinate, ammonium lauryl sulfosuccinate, sodiumdodecylbenzenesulfonate, ammonium laureth sulfate, sodium N-laurylsarcosinate, sodium lauryl sulfate, and combinations thereof. In anotheraspect, the composition comprises one or more buffering agents.

In another embodiment, the present invention includes a nutritionalsupplement comprising: a composition comprising a source of methylsalicylate, an acetyl donor, and L-Arginine, wherein the composition iseffective to produce aspirin-triggered resolvins in the subject withoutthe deleterious effect of the aspirin or aspirin-like compounds in thestomach. In one aspect, the source of methyl salicylate is wintergreenoil or eastern teaberry oil. In another aspect, the source of methylsalicylate is a Gaultheria sp., Betula sp., Spiraea sp. or a Polygalasp. In another aspect, the methyl salicylate is provided in an amountbetween 10 mg and 60 mg. In another aspect, the acetyl donor is providedin an amount between 30 mg and 300 mg. In another aspect, the L-Arginineis provided in an amount between 3 mg and 40 mg. In another aspect, thecomposition is provided in a gelcap, tablet, powder, cream, lotion,liquid, softgel, poultice, suppository, or serum. In another aspect, thecomposition is formulated for oral, sublingual, subcutaneous,percutaneous, intramuscular, nasal, intradermal, pulmonary,intraalveolar, intravaginal, intrarectal, intraperitoneal or intravenousadministration. In another aspect, the composition demonstratesincreased potency and reduced off-target effects when compared toaspirin, aspirin-like products, or taken separately. In another aspect,the acetyl donor is d,l-alpha-Tocopherol Acetate, Vitamin E, or anacetylated vitamin. In another aspect, the composition further comprisesan Omega-3 dietary supplement to the composition. In another aspect, thecomposition further comprises an eicosapentaenoic acid (EPA) to thecomposition. In another aspect, the composition further comprises adocosahexaenoic acid (DHA) to the composition. In another aspect, thecomposition comprises an emulsifier selected from beeswax, ammoniumlauryl sulfate, sodium laureth sulfate, sodium oleyl succinate, ammoniumlauryl sulfosuccinate, sodium dodecylbenzenesulfonate, ammonium laurethsulfate, sodium N-lauryl sarcosinate, sodium lauryl sulfate, andcombinations thereof. In another aspect, the composition comprises oneor more buffering agents.

In another embodiment, the present invention includes a nutritionalsupplement comprising: a source of methyl salicylate, an acetyl donor,and L-Arginine, wherein the composition is effective to produceaspirin-triggered resolvins in the subject without the deleteriouseffect of the aspirin or aspirin-like compounds in the stomach.

In another embodiment, the present invention includes a nutritionalsupplement consists essentially of: a source of methyl salicylate, anacetyl donor, and L-Arginine, wherein the composition is effective toproduce aspirin-triggered resolvins in the subject without thedeleterious effect of the aspirin or aspirin-like compounds in thestomach.

In another embodiment, the present invention includes a nutritionalsupplement consists of: a source of methyl salicylate, an acetyl donor,and L-Arginine, wherein the composition is effective to produceaspirin-triggered resolvins in the subject without the deleteriouseffect of the aspirin or aspirin-like compounds in the stomach.

In another embodiment, the present invention includes a nutritionalsupplement comprising: methyl salicylate, d,l-alpha-Tocopherol Acetate,Vitamin E, or an acetylated vitamin, L-Arginine, eicosapentaenoic acid(EPA) and docosahexaenoic acid (DHA), wherein the supplement iseffective to produce aspirin-triggered resolvins in the subject withoutthe deleterious effect of the aspirin or aspirin-like compounds in thestomach.

In another embodiment, the present invention includes a nutritionalsupplement consisting essentially of: methyl salicylate,d,l-alpha-Tocopherol Acetate, Vitamin E, or an acetylated vitamin,L-Arginine, eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA),wherein the supplement is effective to produce aspirin-triggeredresolvins in the subject without the deleterious effect of the aspirinor aspirin-like compounds in the stomach.

In another embodiment, the present invention includes a nutritionalsupplement consists of: methyl salicylate, d,l-alpha-Tocopherol Acetate,Vitamin E, or an acetylated vitamin, L-Arginine, eicosapentaenoic acid(EPA) and docosahexaenoic acid (DHA), wherein the supplement iseffective to produce aspirin-triggered resolvins in the subject withoutthe deleterious effect of the aspirin or aspirin-like compounds in thestomach.

In another embodiment, the present invention includes a method oftreating a subject in need for pain relief or improved cardiovascularfunction comprising: identifying a subject in need of aspirin oraspirin-like compounds; and providing the subject with a supplementcomprising: a source of methyl salicylate, an acetyl donor, andL-Arginine, wherein the composition is effective to produceaspirin-triggered resolvins in the subject without the deleteriouseffect of the aspirin or aspirin-like compounds in the stomach. In oneaspect, the source of methyl salicylate is wintergreen oil or easternteaberry oil. In another aspect, the source of methyl salicylate is aGaultheria sp., Betula sp., Spiraea sp. or a Polygala sp. In anotheraspect, the methyl salicylate is provided in an amount between 10 mg and60 mg. In another aspect, the acetyl donor is provided in an amountbetween 30 mg and 300 mg. In another aspect, the L-Arginine is providedin an amount between 3 mg and 40 mg. In another aspect, the methodfurther comprises packaging the composition into a gelcap, tablet,powder, cream, lotion, liquid, softgel, poultice, suppository, or serum.In another aspect, the method further comprises formulating thecomposition for oral, sublingual, subcutaneous, percutaneous,intramuscular, nasal, intradermal, pulmonary, intraalveolar,intravaginal, intrarectal, intraperitoneal or intravenousadministration. In another aspect, the composition demonstrates increasepotency and reduced off-target effects when compared to aspirin,aspirin-like products, or taken separately. In another aspect, theacetyl donor is d,l-alpha-Tocopherol Acetate, Vitamin E, or anacetylated vitamin. In another aspect, the method further comprisesadding an Omega-3 dietary supplement the composition.

In another aspect, the method further comprises adding eicosapentaenoicacid (EPA) to the composition. In another aspect, the method furthercomprises adding docosahexaenoic acid (DHA) to the composition. Inanother aspect, the composition comprises an emulsifier selected frombeeswax, ammonium lauryl sulfate, sodium laureth sulfate, sodium oleylsuccinate, ammonium lauryl sulfosuccinate, sodiumdodecylbenzenesulfonate, ammonium laureth sulfate, sodium N-laurylsarcosinate, sodium lauryl sulfate, and combinations thereof. In anotheraspect, the composition comprises one or more buffering agents.

BRIEF DESCRIPTION OF THE DRAWINGS

For a more complete understanding of the features and advantages of thepresent invention, reference is now made to the detailed description ofthe invention along with the accompanying figures and in which:

FIG. 1 shows the basic equation for the generation of aspirin in-situbased on simple principles of organic chemistry of the presentinvention.

FIG. 2 shows a specific example of an in-situ aspirin-generatingreaction.

DETAILED DESCRIPTION OF THE INVENTION

While the making and using of various embodiments of the presentinvention are discussed in detail below, it should be appreciated thatthe present invention provides many applicable inventive concepts thatcan be embodied in a wide variety of specific contexts. The specificembodiments discussed herein are merely illustrative of specific ways tomake and use the invention and do not delimit the scope of theinvention.

To facilitate the understanding of this invention, a number of terms aredefined below. Terms defined herein have meanings as commonly understoodby a person of ordinary skill in the areas relevant to the presentinvention. Terms such as “a”, “an” and “the” are not intended to referto only a singular entity, but include the general class of which aspecific example may be used for illustration. The terminology herein isused to describe specific embodiments of the invention, but their usagedoes not limit the invention, except as outlined in the claims.

The invention described herein relates to a novel method to generateaspirin and aspirin-like compounds in situ from readily available andregulatory compliant materials used in the nutraceutical industry. Thismethodology is incorporated into a fish oil nutritional supplement inorder to produce Aspirin-Triggered Resolvins in the body without the useof aspirin. This new method has the advantages of ease of use,regulatory compliance in the supplement industry, and increasedconversion of Aspirin-Triggered Resolvins due to the proximity effect ofhaving these ingredients all in the same formulation. This is the firstwork to describe a method to create Aspirin-Triggered Resolvins directlyin the body without the use of Aspirin and all in one simpleformulation.

Polyunsaturated fatty acids (PUFAs) from fish oil have demonstrated aprofound ability to maintain human health from a myriad of differentmechanisms. Among the PUFAs in fish oil, eicosapentaenoic acid (EPA) anddocosahexaenoic acid (DHA) are the major bioactive players. Researchover the last 15 years has shown that EPA and DHA are actually prodrugsin the body; their biological activity in vivo is due to the productionof potent anti-inflammatory metabolites called Resolvins (Rv) andProtectins (PD). Different Resolvins and Protectins with varying degreesof anti-inflammatory properties are produced from both EPA and DHA.

Hundreds of clinical studies have been conducted over the years showingthe health benefits of EPA and DHA-containing omega-3 supplements formaintaining cardio health and healthy inflammatory status in the body.Meta-analysis of these studies demonstrates that study outcomes are moreeffective and significant when participants concurrently take alow-regiment dose of acetylsalicylic acid (aspirin). Recent research hasshown that the increased anti-inflammatory properties of taking anomega-3 supplement in the presence of a low dose of aspirin is due tothe acetylation of the cyclooxygenase-2 (COX-2) enzyme which results inthe enzymatic conversion of EPA and DHA into a new class of Resolvinsknown as Aspirin-Triggered Resolvins (AT-Rv) that possess even greateranti-inflammatory properties than Resolvins not triggered by aspirin.

The problem with taking aspirin and an EPA/DHA-containing nutritionalsupplement separately is the chance for off-target effects due to themetabolism of aspirin, EPA, and DHA before they have the chance to reactwith each other in the gut. This invention helps to circumvent thissituation and increase the effectiveness of the conversion of aspirin,EPA, and DHA to Aspirin-Triggered Resolvins.

Dosage Forms.

A dosage unit for use of the composition of the present invention may bemixed together, form ionic or even covalent bonds. The methylsalicylate, d,l-alpha-Tocopherol Acetate, and L-Arginine of the presentinvention may be administered in oral, intravenous (bolus or infusion),intraperitoneal, subcutaneous, or intramuscular form, all using dosageforms well known to those of ordinary skill in the pharmaceutical arts.Depending on the particular location or method of delivery, differentdosage forms, e.g., tablets, capsules, pills, powders, granules,elixirs, tinctures, suspensions, syrups, and emulsions may be used toprovide the methyl salicylate, d,l-alpha-Tocopherol Acetate, andL-Arginine of the present invention to a patient in need of therapy. Themethyl salicylate, d,l-alpha-Tocopherol Acetate, and L-Arginine may alsobe administered as any one of known salt forms.

Methyl salicylate, d,l-alpha-Tocopherol Acetate, and L-Arginine can beadministered in admixture with suitable pharmaceutical salts, buffers,diluents, extenders, excipients and/or carriers (collectively referredto herein as a pharmaceutically acceptable carrier or carrier materials)selected based on the intended form of administration and as consistentwith conventional pharmaceutical practices. Depending on the bestlocation for administration, the methyl salicylate, d,l-alpha-TocopherolAcetate, and L-Arginine may be formulated to provide, e.g., maximumand/or consistent dosing for the particular form for oral, rectal,topical, intravenous injection or parenteral administration. While themethyl salicylate, d,l-alpha-Tocopherol Acetate, and L-Arginine may beadministered alone, it will generally be provided in a stable salt formmixed with a pharmaceutically acceptable carrier. The carrier may besolid or liquid, depending on the type and/or location of administrationselected.

Techniques and compositions for making useful dosage forms using thepresent invention are described in one or more of the followingreferences: Anderson, Philip O.; Knoben, James E.; Troutman, William G,eds., Handbook of Clinical Drug Data, Tenth Edition, McGraw-Hill, 2002;Pratt and Taylor, eds., Principles of Drug Action, Third Edition,Churchill Livingston, N.Y., 1990; Katzung, ed., Basic and ClinicalPharmacology, Ninth Edition, McGraw Hill, 2007; Goodman and Gilman,eds., The Pharmacological Basis of Therapeutics, Tenth Edition, McGrawHill, 2001; Remington's Pharmaceutical Sciences, 20th Ed., LippincottWilliams & Wilkins., 2000; Martindale, The Extra Pharmacopoeia,Thirty-Second Edition (The Pharmaceutical Press, London, 1999); all ofwhich are incorporated by reference, and the like, relevant portionsincorporated herein by reference.

For example, the methyl salicylate, d,l-alpha-Tocopherol Acetate, andL-Arginine may be included in a tablet. Tablets may contain, e.g.,suitable binders, lubricants, disintegrating agents, coloring agents,flavoring agents, flow-inducing agents and/or melting agents. Forexample, oral administration may be in a dosage unit form of a tablet,gelcap, caplet or capsule, the active drug component being combined witha non-toxic, pharmaceutically acceptable, inert carrier such as lactose,gelatin, agar, starch, sucrose, glucose, methyl cellulose, magnesiumstearate, dicalcium phosphate, calcium sulfate, mannitol,sorbitol,mixtures thereof, and the like. Suitable binders for use withthe present invention include: starch, gelatin, natural sugars (e.g.,glucose or beta-lactose), corn sweeteners, natural and synthetic gums(e.g., acacia, tragacanth or sodium alginate), carboxymethylcellulose,polyethylene glycol, waxes, and the like. Lubricants for use with theinvention may include: sodium oleate, sodium stearate, magnesiumstearate, sodium benzoate, sodium acetate, sodium chloride, mixturesthereof, and the like. Disintegrators may include: starch, methylcellulose, agar, bentonite, xanthan gum, mixtures thereof, and the like.

Methyl salicylate, d,l-alpha-Tocopherol Acetate, and L-Arginine may beadministered in the form of liposome delivery systems, e.g., smallunilamellar vesicles, large unilamallar vesicles, and multilamellarvesicles, whether charged or uncharged. Liposomes may include one ormore: phospholipids (e.g., cholesterol), stearylamine and/orphosphatidylcholines, mixtures thereof, and the like.

Methyl salicylate, d,l-alpha-Tocopherol Acetate, and L-Arginine may alsobe coupled to one or more soluble, biodegradable, bioacceptable polymersas drug carriers or as a prodrug. Such polymers may include:polyvinylpyrrolidone, pyran copolymer,polyhydroxylpropylmethacrylamide-phenol,polyhydroxyethylasparta-midephenol, or polyethyleneoxide-polylysinesubstituted with palmitoyl residues, mixtures thereof, and the like.Furthermore, the methyl salicylate, d,l-alpha-Tocopherol Acetate, andL-Arginine may be coupled one or more biodegradable polymers to achievecontrolled release of the methyl salicylate, d,l-alpha-TocopherolAcetate, and L-Arginine, biodegradable polymers for use with the presentinvention include: polylactic acid, polyglycolic acid, copolymers ofpolylactic and polyglycolic acid, polyepsilon caprolactone, polyhydroxybutyric acid, polyorthoesters, polyacetals, polydihydropyrans,polycyanoacylates, and crosslinked or amphipathic block copolymers ofhydrogels, mixtures thereof, and the like.

In one embodiment, gelatin capsules (gelcaps) may include the methylsalicylate, d,l-alpha-Tocopherol Acetate, and L-Arginine and powderedcarriers, such as lactose, starch, cellulose derivatives, magnesiumstearate, stearic acid, and the like. Like diluents may be used to makecompressed tablets. Both tablets and capsules may be manufactured asimmediate-release, mixed-release or sustained-release formulations toprovide for a range of release of medication over a period of minutes tohours. Compressed tablets may be sugar coated or film coated to mask anyunpleasant taste and protect the tablet from the atmosphere. An entericcoating may be used to provide selective disintegration in, e.g., thegastrointestinal tract.

For oral administration in a liquid dosage form, the oral drugcomponents may be combined with any oral, non-toxic, pharmaceuticallyacceptable inert carrier such as ethanol, glycerol, water, and the like.Examples of suitable liquid dosage forms include solutions orsuspensions in water, pharmaceutically acceptable fats and oils,alcohols or other organic solvents, including esters, emulsions, syrupsor elixirs, suspensions, solutions and/or suspensions reconstituted fromnon-effervescent granules and effervescent preparations reconstitutedfrom effervescent granules. Such liquid dosage forms may contain, forexample, suitable solvents, preservatives, emulsifying agents,suspending agents, diluents, sweeteners, thickeners, and melting agents,mixtures thereof, and the like.

Liquid dosage forms for oral administration may also include coloringand flavoring agents that increase patient acceptance and thereforecompliance with a dosing regimen. In general, water, a suitable oil,saline, aqueous dextrose (e.g., glucose, lactose and related sugarsolutions) and glycols (e.g., propylene glycol or polyethylene glycols)may be used as suitable carriers for parenteral solutions. Solutions forparenteral administration include generally, a water soluble salt of theactive ingredient, suitable stabilizing agents, and if necessary,buffering salts. Antioxidizing agents such as sodium bisulfate, sodiumsulfite and/or ascorbic acid, either alone or in combination, aresuitable stabilizing agents. Citric acid and its salts and sodium EDTAmay also be included to increase stability. In addition, parenteralsolutions may include pharmaceutically acceptable preservatives, e.g.,benzalkonium chloride, methyl- or propyl-paraben, and/or chlorobutanol.Suitable pharmaceutical carriers are described in Remington'sPharmaceutical Sciences, Mack Publishing Company, a standard referencetext in this field, relevant portions incorporated herein by reference.

For direct delivery to the nasal passages, sinuses, mouth, throat,esophagus, trachea, lungs and alveoli, the methyl salicylate,d,l-alpha-Tocopherol Acetate, and L-Arginine may also be delivered as anintranasal form via use of a suitable intranasal vehicle. For dermal andtransdermal delivery, the methyl salicylate, d,l-alpha-TocopherolAcetate, and L-Arginine may be delivered using lotions, creams, oils,elixirs, serums, transdermal skin patches and the like, as are wellknown to those of ordinary skill in that art. Parenteral and intravenousforms may also include pharmaceutically acceptable salts and/or mineralsand other materials to make them compatible with the type of injectionor delivery system chosen, e.g., a buffered, isotonic solution. Examplesof useful pharmaceutical dosage forms for administration of methylsalicylate, d,l-alpha-Tocopherol Acetate, and L-Arginine may include thefollowing forms.

Capsules. Capsules may be prepared by filling standard two-piece hardgelatin capsules each with 10 to 500 milligrams of powdered activeingredient, 5 to 150 milligrams of lactose, 5 to 50 milligrams ofcellulose and 6 milligrams magnesium stearate.

Soft Gelatin Capsules. A mixture of active ingredient is dissolved in adigestible oil such as soybean oil, cottonseed oil or olive oil. Theactive ingredient is prepared and injected by using a positivedisplacement pump into gelatin to form soft gelatin capsules containing,e.g., 100-500 milligrams of the active ingredient. The capsules arewashed and dried.

Tablets. A large number of tablets are prepared by conventionalprocedures so that the dosage unit was 100-500 milligrams of activeingredient, 0.2 milligrams of colloidal silicon dioxide, 5 milligrams ofmagnesium stearate, 50-275 milligrams of microcrystalline cellulose, 11milligrams of starch and 98.8 milligrams of lactose. Appropriatecoatings may be applied to increase palatability or delay absorption.

To provide an effervescent tablet with the methyl salicylate,d,l-alpha-Tocopherol Acetate, and L-Arginine, appropriate amounts of,e.g., monosodium citrate and sodium bicarbonate, are blended togetherand then roller compacted, in the absence of water, to form flakes thatare then crushed to give granulates. The granulates are then combinedwith the active ingredient, drug and/or salt thereof, conventionalbeading or filling agents and, optionally, sweeteners, flavors andlubricants.

Injectable solution. A parenteral composition suitable foradministration by injection is prepared by stirring 1.5% by weight ofactive ingredient in deionized water and mixed with, e.g., up to 10% byvolume propylene glycol and water. The solution is made isotonic withsodium chloride and sterilized using, e.g., ultrafiltration.

Suspension. An aqueous suspension is prepared for oral administration sothat each 5 ml contain 100 mg of finely divided active ingredient, 200mg of sodium carboxymethyl cellulose, 5 mg of sodium benzoate, 1.0 g ofsorbitol solution, U.S.P., and 0.025 ml of vanillin.

For mini-tablets, the active ingredient is compressed into a hardness inthe range 6 to 12 Kp. The hardness of the final tablets is influenced bythe linear roller compaction strength used in preparing the granules orgranulates, which are influenced by the particle size of, e.g., themonosodium hydrogen carbonate and sodium hydrogen carbonate.

For smaller particle sizes, a linear roller compaction strength of about15 to 20 KN/cm may be used.

Examples of suitable liquid dosage forms include solutions orsuspensions in water, pharmaceutically acceptable fats and oils,alcohols or other organic solvents, including esters, emulsions, syrupsor elixirs, suspensions, solutions and/or suspensions reconstituted fromnon-effervescent granules and effervescent preparations reconstitutedfrom effervescent granules. Such liquid dosage forms may contain, forexample, suitable solvents, preservatives, emulsifying agents,suspending agents, diluents, sweeteners, thickeners, and melting agents.Oral dosage forms optionally contain flavorants and coloring agents.Parenteral and intravenous forms may also include minerals and othermaterials to make them compatible with the type of injection or deliverysystem chosen.

The composition of methyl salicylate, d,l-alpha-Tocopherol Acetate, andL-Arginine may be formulated for release that is immediate, rapid,extended, bi-phasic, etc. By “immediate release” is meant a release ofan active agent to an environment over a period of seconds to no morethan about 30 minutes once release has begun and release begins withinno more than about 2 minutes after administration. An immediate releasedoes not exhibit a significant delay in the release of drug. By “rapidrelease” is meant a release of an active agent to an environment over aperiod of 1-59 minutes or 0.1 minute to three hours once release hasbegun and release can begin within a few minutes after administration orafter expiration of a delay period (lag time) after administration. Asused herein, the term “extended release” profile assumes the definitionas widely recognized in the art of pharmaceutical sciences. An extendedrelease dosage form will release drug at substantially constant rateover an extended period of time or a substantially constant amount ofdrug will be released incrementally over an extended period of time. Anextended release tablet generally effects at least a two-fold reductionin dosing frequency as compared to the drug presented in a conventionaldosage form (e.g., a solution or rapid releasing conventional soliddosage forms). By “controlled release” is meant a release of an activeagent to an environment over a period of about eight hours up to about12 hours, 16 hours, 18 hours, 20 hours, a day, or more than a day. By“sustained release” is meant an extended release of an active agent tomaintain a constant drug level in the blood or target tissue of asubject to which the device is administered. The term “controlledrelease”, as regards to drug release, includes the terms “extendedrelease”, “prolonged release”, “sustained release”, or “slow release”,as these terms are used in the pharmaceutical sciences. An controlledrelease can begin within a few minutes after administration or afterexpiration of a delay period (lag time) after administration.

A slow release dosage form is one that provides a slow rate of releaseof drug so that drug is released slowly and approximately continuouslyover a period of 3 hours, 6 hours, 12 hours, 18 hours, a day, 2 or moredays, a week, or 2 or more weeks, for example.

A timed-release dosage form is one that begins to release drug after apredetermined period of time as measured from the moment of initialexposure to the environment of use.

A targeted release dosage form generally refers to an oral dosage formthat designed to deliver drug to a particular portion of thegastrointestinal tract of a subject. An exemplary targeted dosage formis an enteric dosage form that delivers a drug into the middle to lowerintestinal tract but not into the stomach or mouth of the subject. Othertargeted dosage forms can deliver to other sections of thegastrointestinal tract such as the stomach, jejunum, ileum, duodenum,cecum, large intestine, small intestine, colon, or rectum.

By “delayed release” it is meant that initial release of drug occursafter expiration of an approximate delay (or lag) period. For example,if release of drug from an extended release composition is delayed twohours, then release of drug from begins at about two hours afteradministration of the composition, or dosage form, to a subject. Ingeneral, a delayed release is opposite an immediate release, whereinrelease of drug begins after no more than a few minutes afteradministration. Accordingly, the drug release profile from a particularcomposition can be a delayed-extended release or a delayed-rapidrelease. A “delayed-extended” release profile is one wherein extendedrelease of drug begins after expiration of an initial delay period. A“delayed-rapid” release profile is one wherein rapid release of drugbegins after expiration of an initial delay period.

A pulsatile release dosage form is one that provides pulses of highactive ingredient concentration, interspersed with low concentrationtroughs. A pulsatile profile containing two peaks may be described as“bimodal” or “biphasic”.

A pseudo-first order release profile is one that approximates a firstorder release profile. A first order release profile characterizes therelease profile of a dosage form that releases a constant percentage ofan initial drug charge per unit time.

A pseudo-zero order release profile is one that approximates azero-order release profile. A zero-order release profile characterizesthe release profile of a dosage form that releases a constant amount ofdrug per unit time.

The resulting product may also be formulated to exhibit enhanceddissolution rate of a formulated poorly water soluble drug.

The present invention includes, but is not limited to, the following:(1) a method for producing aspirin both in vivo and in situ through theuse of simple commercially available starting materials. (2) A methodusing wintergreen oil (a source of Methyl Salicylate),d,l-alpha-Tocopherol Acetate, and L-Arginine. (3) Coupling this in situaspirin-generating method with EPA/DHA-containing fish oil to produceAspirin-Triggered Resolvins in the body all in a single delivery form,and without the need for taking aspirin separately. (4) Thisaspirin-generating methodology is used in this case with a EPA/DHAcontaining fish oil but could also be used either by itself or withother compounds known to have a beneficial effect when take concurrentlywith aspirin.

In addition to wintergreen oil, vitamin E acetate, and L-arginine, otherin ingredients are EPA, and/or DHA. The aspirin-generation compositionand methods described herein can be used separately in a nutritionalsupplement to generate aspirin in the body. This composition and methodstaught herein use common nutritional ingredients approved by globalregulatory agencies for use in nutritional/dietary supplements. Thecomposition and methods trigger the in situ generation of aspirin in thesame formulation as EPA and DHA might increase potency and reduceoff-target effects that are generated when taken separately. Thisformulation can be made in a soft gel delivery form with beeswax used asan emulsifier in order to increase solubility and material uniformity.The composition and methods for in situ aspirin-formation could also beapplied in powder, tablet, cream, lotion, and liquid delivery forms. Thecomposition and methods for in situ aspirin-generation can also beapplied to any application wherein the generation of aspirin would bebeneficial to human health.

FIG. 1 shows the basic equation for the generation of aspirin in situbased on simple principles of organic chemistry is shown in. The firststep in this process is selection of a base that can deprotonate thephenolic hydrogen of methyl salicylate (which is the major component of,e.g., wintergreen oil). The pKa of this hydrogen is going to be lowerthan the usual number of 10 for phenolic alcohols due to the additionalstabilization of the ester group on the aromatic ring. As such, arelatively weak base (like the amine of L-arginine) should be able todeprotonate the phenolic hydrogen to the alkoxide. The next step in thereaction is to add a natural compound that is an acetyl donor andregulatory compliant in the nutraceutical industry. Acetyl derivativesof common vitamins (like d,l-alpha-tocopherol acetate) are one choice.The mechanism of this reaction involves deprotonation of the phenolichydrogen by the base to form the alkoxide. Next, the nucleophilicalkoxide would attack the carbonyl carbon of the acetyl group on thedonor molecule, which, upon formation and collapse of the tetrahedralintermediate, provides methyl acetylsalicylate and an alcohol as abyproduct. The ester group of methyl acetylsalicylate is cleaved in vivoby many esterase enzymes in the body to yield aspirin.

FIG. 2 shows one example of specific ingredients for thisaspirin-generating reaction. Wintergreen oil is mostly comprised ofmethyl salicylate and is one starting material of choice due to itsallowed regulatory status, cheap price, plentiful supply chain, andstriking similarity in chemical structure to aspirin. The base used inthe example below is the free base form of L-arginine—which is alsowidely available and used in the nutritional supplement industry.Vitamin E acetate is used as the acetyl donor due to the labile natureof the acetyl group and also due to the wide acceptability of Vitamin Ein the nutraceutical industry. The benefit of using L-Arginine andVitamin E in this reaction is the additional health functional benefitsof these ingredients in addition to the in situ generation of aspirin.

As such, the present invention includes a novel method for the in situformation of aspirin or aspirin-like products in the body from readilyavailable starting materials in the nutraceutical industry. This in situaspirin or aspirin-like product generation method can been added to aOmega-3 dietary supplement in order to convert EPA and DHA in fish oilinto a class of compounds called Aspirin-Triggered Resolvins thatpossess potent anti-inflammatory activity and are key class of compoundsresponsible for the clinically-validated biological activity of omega-3supplementation.

It is contemplated that any embodiment discussed in this specificationcan be implemented with respect to any method, kit, reagent, orcomposition of the invention, and vice versa. Furthermore, compositionsof the invention can be used to achieve methods of the invention.

It will be understood that particular embodiments described herein areshown by way of illustration and not as limitations of the invention.The principal features of this invention can be employed in variousembodiments without departing from the scope of the invention. Thoseskilled in the art will recognize, or be able to ascertain using no morethan routine experimentation, numerous equivalents to the specificprocedures described herein. Such equivalents are considered to bewithin the scope of this invention and are covered by the claims.

All publications and patent applications mentioned in the specificationare indicative of the level of skill of those skilled in the art towhich this invention pertains. All publications and patent applicationsare herein incorporated by reference to the same extent as if eachindividual publication or patent application was specifically andindividually indicated to be incorporated by reference.

The use of the word “a” or “an” when used in conjunction with the term“comprising” in the claims and/or the specification may mean “one,” butit is also consistent with the meaning of “one or more,” “at least one,”and “one or more than one.” The use of the term “or” in the claims isused to mean “and/or” unless explicitly indicated to refer toalternatives only or the alternatives are mutually exclusive, althoughthe disclosure supports a definition that refers to only alternativesand “and/or.” Throughout this application, the term “about” is used toindicate that a value includes the inherent variation of error for thedevice, the method being employed to determine the value, or thevariation that exists among the study subjects.

As used in this specification and claim(s), the words “comprising” (andany form of comprising, such as “comprise” and “comprises”), “having”(and any form of having, such as “have” and “has”), “including” (and anyform of including, such as “includes” and “include”) or “containing”(and any form of containing, such as “contains” and “contain”) areinclusive or open-ended and do not exclude additional, unrecitedelements or method steps. In embodiments of any of the compositions andmethods provided herein, “comprising” may be replaced with “consistingessentially of” or “consisting of”. As used herein, the phrase“consisting essentially of” requires the specified integer(s) or stepsas well as those that do not materially affect the character or functionof the claimed invention. As used herein, the term “consisting” is usedto indicate the presence of the recited integer (e.g., a feature, anelement, a characteristic, a property, a method/process step or alimitation) or group of integers (e.g., feature(s), element(s),characteristic(s), property(ies), method/process steps or limitation(s))only.

The term “or combinations thereof” as used herein refers to allpermutations and combinations of the listed items preceding the term.For example, “A, B, C, or combinations thereof” is intended to includeat least one of: A, B, C, AB, AC, BC, or ABC, and if order is importantin a particular context, also BA, CA, CB, CBA, BCA, ACB, BAC, or CAB.Continuing with this example, expressly included are combinations thatcontain repeats of one or more item or term, such as BB, AAA, AB, BBC,AAABCCCC, CBBAAA, CABABB, and so forth. The skilled artisan willunderstand that typically there is no limit on the number of items orterms in any combination, unless otherwise apparent from the context.

As used herein, words of approximation such as, without limitation,“about”, “substantial” or “substantially” refers to a condition thatwhen so modified is understood to not necessarily be absolute or perfectbut would be considered close enough to those of ordinary skill in theart to warrant designating the condition as being present. The extent towhich the description may vary will depend on how great a change can beinstituted and still have one of ordinary skilled in the art recognizethe modified feature as still having the required characteristics andcapabilities of the unmodified feature. In general, but subject to thepreceding discussion, a numerical value herein that is modified by aword of approximation such as “about” may vary from the stated value byat least ±1, 2, 3, 4, 5, 6, 7, 10, 12 or 15%.

All of the compositions and/or methods disclosed and claimed herein canbe made and executed without undue experimentation in light of thepresent disclosure. While the compositions and methods of this inventionhave been described in terms of preferred embodiments, it will beapparent to those of skill in the art that variations may be applied tothe compositions and/or methods and in the steps or in the sequence ofsteps of the method described herein without departing from the concept,spirit and scope of the invention. All such similar substitutes andmodifications apparent to those skilled in the art are deemed to bewithin the spirit, scope and concept of the invention as defined by theappended claims.

REFERENCES

-   Groeger, et al., Cyclooxygenase-2 generates anti-inflammatory    mediators from omega-3 fatty acids, NATURE CHEMICAL BIOLOGY, Vol. 6,    June 2010, 433-441.-   Serhan, et al., Resolvins: A Family of Bioactive Products of Omega-3    Fatty Acid Transformation Circuits Initiated by Aspirin Treatment    that Counter Proinflammation Signals, J. Exp. Med., Volume 196,    Number 8, Oct. 21, 2002 1025-1037.-   Dalli, et al., Resolvin D3 and Aspirin-Triggered Resolvin D3 Are    Potent Immunoresolvents, Chemistry & Biology 20, 188-201, Feb. 21,    2013.-   Serhan, et al., Novel Proresolving Aspirin-Triggered DHA Pathway,    Chemistry & Biology 18, 976-987, Aug. 26, 2011.-   Serhan, et al., Novel Functional Sets of Lipid-derived Mediators    with Antiinflammatory Actions Generated from Omega-3 Fatty Acids via    Cyclooxygenase 2-Nonsteroidal Antiinflammatory Drugs and    Transcellular Processing, J. Exp. Med., Volume 192, Number 8, Oct.    16, 2000 1197-1204.-   Chen, C., COX-2's new role in inflammation, NATURE CHEMICAL BIOLOGY,    Vol 6, June 2010, 401-402.-   Serhan, C., Novel Pro-Resolving Lipid Mediators in Inflammation Are    Leads for Resolution Physiology, Nature. 2014 June 5; 510(7503):    92-101.-   Xu, et al., Resolvins RvE1 and RvD1 attenuate inflammatory pain via    central and peripheral actions, NATURE MEDICINE, Volume 16, Number    5, May 2010, 592-598.-   Morris, et al., Effects of Low-Dose Aspirin on Acute Inflammatory    Responses in Humans, J Immunol 2009; 183:2089-2096; 13 Jul. 2009.-   Serhan, C., Novel N w 3-derived local mediators in anti-inflammation    and resolution, Pharmacology & Therapeutics 105 (2005) 7-21.-   Schwab, et al., Resolvin El and protectin D1 activate    inflammation-resolution programmes, Nature, Vol 447, 14 Jun. 2007,    869-875.-   Ariota, et al., Stereochemical assignment, anti-inflammatory    properties, and receptor for the omega-3 lipid mediator resolvin    El, J. Exp. Med., Vol. 201, No. 5, Mar. 7, 2005 713-722.-   Buckley, et al., Proresolving Lipid Mediators and Mechanisms in the    Resolution of Acute Inflammation, Immunity 40, Mar. 20, 2014,    315-327.-   Ogata, et al., Effects of aspirin-triggered resolvin D1 on    peripheral blood mononuclear cells from patients with Chagas' heart    disease, European Journal of Pharmacology 777 (2016) 26-32.-   Lima-Garcia, et al., The precursor of resolvin D series and    aspirin-triggered resolvin D1 display anti-hyperalgesic properties    in adjuvant-induced arthritis in rats, British Journal of    Pharmacology (2011) 164 278-293.-   Kohli, Resolvins and protectins: mediating solutions to    inflammation, British Journal of Pharmacology (2009), 158, 960-971.-   Prescott and McKay, Aspirin-triggered lipoxin enhances macrophage    phagocytosis of bacteria while inhibiting inflammatory cytokine    production, Am J Physiol Gastrointest Liver Physiol 301: G487-G497,    2011.-   Skarke, et al., Bioactive products formed in humans from fish    oils, J. of Lipid Research, Sep. 29, 2015, pages 1-42.-   Sok, et al., Aspirin-triggered resolvin D1-modified materials    promote the accumulation of pro-regenerative immune cell subsets and    enhance vascular remodeling, Acta Biomaterialia (2017), 1-34.

What is claimed is:
 1. A method of producing aspirin in situ, the method comprising: identifying a subject in need of aspirin or aspirin-like compounds; and providing the subject with a composition comprising: a source of methyl salicylate, an acetyl donor, and L-Arginine, wherein the composition is effective to produce aspirin-triggered resolvins in the subject without the deleterious effect of the aspirin or aspirin-like compounds in the stomach.
 2. The method of claim 1, wherein the source of methyl salicylate is wintergreen oil or eastern teaberry oil.
 3. The method of claim 1, wherein the source of methyl salicylate is a Gaultheria sp., Betula sp., Spiraea sp. or a Polygala sp.
 4. The method of claim 1, wherein at least one of: the methyl salicylate is provided in an amount between 10 mg and 60 mg, the acetyl donor is provided in an amount between 30 mg and 300 mg, or the L-Arginine is provided in an amount between 3 mg and 40 mg.
 5. The method of claim 1, further comprising packaging the composition into a gelcap, tablet, powder, cream, lotion, liquid, softgel, poultice, suppository, or serum.
 6. The method of claim 1, further comprising formulating the composition for oral, sublingual, subcutaneous, percutaneous, intramuscular, nasal, intradermal, pulmonary, intraalveolar, intravaginal, intrarectal, intraperitoneal or intravenous administration.
 7. The method of claim 1, wherein the composition demonstrates increased potency and reduced off-target effects when compared to aspirin or aspirin-like products taken separately.
 8. The method of claim 1, wherein the acetyl donor is d,l-alpha-Tocopherol Acetate, Vitamin E, or an acetylated vitamin.
 9. The method of claim 1, further comprising adding at least one of: an Omega-3 dietary supplement, eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), or one or more buffering agents to the composition.
 10. The method of claim 1, wherein the composition comprises an emulsifier selected from beeswax, ammonium lauryl sulfate, sodium laureth sulfate, sodium oleyl succinate, ammonium lauryl sulfosuccinate, sodium dodecylbenzenesulfonate, ammonium laureth sulfate, sodium N-lauryl sarcosinate, sodium lauryl sulfate, and combinations thereof.
 11. A nutritional supplement comprising: a composition comprising a source of a methyl salicylate, an acetyl donor, and L-Arginine, wherein the composition is effective to produce aspirin-triggered resolvins in the subject without the deleterious effect of the aspirin or aspirin-like compounds in the stomach.
 12. A method of treating a subject in need for pain relief or improved cardiovascular function comprising: identifying a subject in need of aspirin or aspirin-like compounds; and providing the subject with a composition comprising a source of methyl salicylate selected from an oil from a Gaultheria sp Betula sp., Spiraea sp. or a Polygala sp., an acetyl donor selected from d,l-alpha-Tocopherol Acetate, or an acetylated vitamin, and L-Arginine, wherein the composition is effective to produce aspirin-triggered resolvins in the subject without the deleterious effect of the aspirin or aspirin-like compounds in the stomach of the subject.
 13. The method of claim 12, wherein the source of methyl salicylate is wintergreen oil or eastern teaberry oil.
 14. The method of claim 12, wherein the at least one of: the methyl salicylate is provided in an amount between 10 mg and 60 mg, the acetyl donor is provided in an amount between 30 mg and 300 mg, or the L-Arginine is provided in an amount between 3 mg and 40 mg.
 15. The method of claim 12, further comprising packaging the composition into a gelcap, tablet, powder, cream, lotion, liquid, softgel, poultice, suppository, or serum.
 16. The method of claim 12, further comprising formulating the composition for oral, sublingual, subcutaneous, percutaneous, intramuscular, nasal, intradermal, pulmonary, intraalveolar, intravaginal, intrarectal, intraperitoneal or intravenous administration.
 17. The method of claim 12, wherein the composition demonstrates increase potency and reduced off-target effects when compared to aspirin or aspirin-like products taken separately.
 18. The method of claim 12, further comprising adding at least one of: an Omega-3 dietary supplement, an eicosapentaenoic acid (EPA), an docosahexaenoic acid (DHA), or one or more buffering agents an Omega-3 dietary supplement the composition.
 19. The method of claim 12, wherein the composition further comprises an emulsifier selected from beeswax, ammonium lauryl sulfate, sodium laureth sulfate, sodium oleyl succinate, ammonium lauryl sulfosuccinate, sodium dodecylbenzenesulfonate, ammonium laureth sulfate, sodium N-lauryl sarcosinate, sodium lauryl sulfate, and combinations thereof. 